In recent years regenerative medical techniques have been in the limelight. In regenerative medical techniques, disorders that the natural, inherent regenerative-healing ability of the human body cannot cure can be cured by regenerating organs and such using artificial proliferation of autologous cells, and then surgically conjugating these at the site of the lesions. Such cures have been successful in a wide variety of fields.
Transplantation of oligodendroglia (oligodendrocytes) (see Non-patent Documents 1 to 3), or myelin-forming cells, such as Schwann cells (see Non-patent Documents 4, 2, and 5) or olfactory ensheathing cells (see Non-patent Documents 6 to 8), can elicit remyelination in animal models and electrophysiological function may be recovered (see Non-patent Documents 9 and 5). It is not impossible to prepare such cells from patients or other persons for use in cell therapy; however, it is problematic since tissue material must be collected from either the brain or nerves.
Neural progenitor cells or stem cells derived from the brain have the ability to self-proliferate, and are known to differentiate into neurons and glial cells of various lineages (see Non-patent Documents 10 to 13). Upon transplantation into newborn mouse brains, human neural stem cells collected from fetal tissues differentiate into neurons and astrocytes (see Non-patent Documents 14 to 16), and can remyelinate axons (Non-patent Document 17). There have been reports of the remyelination and recovery of impulse conduction when neural progenitor cells derived from adult human brains are transplanted into demyelinated rodent spinal cords (Non-patent Document 18).
These studies have evoked great interest since they indicate the possibility of applying the above-mentioned cells in reparative strategies for neurological diseases (see Non-patent Documents 18, 14 to 16, and 19).
Recent studies have revealed that neural stem cells can produce hematopoietic cells in vivo, indicating that neural progenitor cells are not limited to nervous system cell lineages (see Non-patent Document 20). Further, when bone marrow interstitial cells are injected into newborn mouse lateral ventricles, they differentiate, to a very small extent, into cells expressing astrocyte markers (see Non-patent Document 21). Under appropriate cell culture conditions bone marrow interstitial cells are reported to produce a very small number of cells that express nervous system cell markers in vitro; however, it is unclear whether these cells are useful for neural regeneration (see Non-patent Document 22).
The present inventors have previously extracted and cultured nervous system cells (neural stem cells, neural progenitor cells) from adult human brains, and established some cell lines.
By studying the functions of these cells, the inventors discovered that neural stem cells are pluripotent and can self-reproduce (see Non-patent Document 18). Specifically, single-cell expansion of neural progenitor (stem) cells obtained from adult human brains was conducted to establish cell lines; the established cells were then subjected to in vitro clonal analysis. The results demonstrated that the cell lines were pluripotent (namely, had the ability to differentiate into neurons, astroglia (or astrocytes), and oligodendroglia (i.e., oligodendrocytes)) and had self-reproducing ability (namely, proliferation potency). Thus, these cells were confirmed to possess the characteristics of neural stem cells.
Transplantation of cultured neural stem cells, which were extracted from small amounts of neural tissue collected from the cerebrum of an individual, into a lesion of the brain or spinal cord of the individual, seems to be a widely applicable therapeutic method in autotransplantation therapy. However, although it doesn't cause symptoms of neurological deficiency, collecting tissues that contain neural stem cells from the cerebrum is not easy. Thus, considering the current need to establish therapeutic methods for various complicated diseases of the nervous system, it is crucial to establish safer and simpler methods for autotransplantation therapy. Thus, to obtain donor cells, the present inventors have developed techniques for collecting mononuclear cell fractions and the like from bone marrow cells, cord blood cells, or fetal liver cells, which is simpler than collecting neural stem cells (see Patent Document 1). Specifically, the present inventors have shown that mononuclear cell fractions prepared from bone marrow cells have the ability to differentiate into nervous system cells. They also have shown that cell fractions containing mesodermal stem cells (mesenchymal stem cells), stromal cells, and AC133-positive cells, which were separated from the mononuclear cell fraction, also had the ability to differentiate into nervous system cells.
Cranial nerve diseases can be treated by directly administering an affected part in the brain with cells that have the ability to differentiate into the above-mentioned nervous system cells. This technique, however, is very complicated and dangerous. There is therefore much demand for the development of simple and safe methods and agents for treating cranial nerve diseases.
Mesenchymal stem cells (MSCs) are thought to represent a very small proportion of cells in the mononuclear population of bone marrow. These cells will grow to confluency in appropriate culture conditions as flattened fibroblast-like cells, and have been suggested to differentiate into bone, cartilage, cardiac myocytes and neurons and glia both in vitro and in vivo. MSCs prepared from human bone marrow (BMSCs) have been used in clinical studies for metachromatic leukodystrophy, Hurler syndrome, myeloablative therapy for breast cancer [11], graft-versus-host disease, and stroke.
Human mesenchymal precursor cells found in the blood of normal subjects proliferated in culture with an adherent-spread morphology, and displayed cytoskeletal, cytoplasmic and surface markers (CD34−, CD45−, and CD105+) of mesenchymal precursors. These cells had a capacity for differentiation into fibroblast, osteoblast, and adipocyte lineages. A canine CD34− fibroblast-like cell in the peripheral blood showed mesenchymal stem cell characteristics. Because peripheral blood is readily accessible, stem cells isolated from blood may be a good candidate for a cell therapy.
Transplantation of mesenchymal stem cells derived from bone marrow (BMSCs) after ischemia onset can reduce infarction size and improve functional outcome in rodent cerebral ischemia models. While intravenous injection of BMSCs reduces infarction size and improves functional outcome in a rat stroke model, the therapeutic benefit of MSC-like multipotent precursor cells derived from peripheral blood (PMSCs) transplantation in cerebral ischemia is still uncertain.
Although the potential of MSCs in peripheral blood (PMSCs) has been studied, it is not previously been known whether peripheral blood-derived plastic-adherent stem/precursor cells (PMSCs) can differentiate into a neural lineage or provide a therapeutic benefit for victims of stroke.    [Patent Document] WO 02/00849    [Non-patent Document 1] Archer D R, et al. 1994. Exp Neurol 125:268-77.    [Non-patent Document 2] Blakemore W F, Crang A J. 1988. Dev Neurosci 10:1-11.    [Non-patent Document 3] Gumpel M, et al. 1987. Ann New York Acad Sci 495:71-85.    [Non-patent Document 4] Blakemore W F. 1977. Nature 266:68-9.    [Non-patent Document 5] Honmou O, et al. 1996. J Neurosci 16:3199-208.    [Non-patent Document 6] Franklin R J, et al. 1996. Glia 17:217-24.    [Non-patent Document 7] Imaizumi T, et al. 1998. J Neurosci 18(16):6176-6185.    [Non-patent Document 8] Kato T, et al. 2000. Glia 30:209-218.    [Non-patent Document 9] Utzschneider D A, et al. 1994. Proc Natl Acad Sci USA 91:53-7.    [Non-patent Document 10] Gage F H, et al. 1995. Proc Natl Acad Sci USA 92:11879-83.    [Non-patent Document 11] Lois C, Alvarez-Buylla A. 1993. Proc Natl Acad Sci USA 90:2074-7.    [Non-patent Document 12] Morshead C M, et al. 1994. Neuron 13:1071-82.    [Non-patent Document 13] Reynolds B A, Weiss S. 1992. Science 255:1707-10.    [Non-patent Document 14] Chalmers-Redman R M, et al. 1997. Neurosci 76:1121-8.    [Non-patent Document 15] Moyer M P, et al. 1997. Transplant Proc 29:2040-1.    [Non-patent Document 16] Svendsen C N, et al. 1997. Exp Neurol 148:135-46.    [Non-patent Document 17] Flax J D, et al. 1998. Nat Biotechnol 16:1033-9.    [Non-patent Document 18] Akiyama Y, et al. 2001. Exp Neurol.    [Non-patent Document 19] Yandava B D, et al. 1999. Proc Natl Acad Sci USA 96:7029-34.    [Non-patent Document 20] Bjornson C R, et al. 1999. Science 283:534-7.    [Non-patent Document 21] Kopen G C, et al. Proc Natl Acad Sci USA 96:10711-6.    [Non-patent Document 22] Woodbury D, et al. 2000. J Neurosci Res 61:364-70.    [Non-patent document 23] Friedenstein A J. Precursor cells of mechanocytes. Int Rev Cytol 1976; 47:327-359.    [Non-patent document 24] Majumdar M K, Thiede M A, Mosca J D et al. Phenotypic and functional comparison of cultures of marrow-derived mesenchymal stem cells (MSCs) and stromal cells. J Cell Physiol 1998; 176:57-66.    [Non-patent document 25] Kobune M, Kawano Y, Ito Y et al. Telomerized human multipotent mesenchymal cells can differentiate into hematopoietic and cobblestone area-supporting cells. Exp Hematol 2003; 31:715-722.    [Non-patent document 26] Toma C, Pittenger M F, Cahill K S et al. Human mesenchymal stem cells differentiate to a cardiomyocyte phenotype in the adult murine heart. Circulation 2002; 105:93-98.    [Non-patent document 27] Prockop D J. Marrow stromal cells as stem cells for nonhematopoietic tissues. Science 1997; 276:71-74.    [Non-patent document 28] Woodbury D, Schwarz E J, Prockop D J et al. Adult rat and human bone marrow stromal cells differentiate into neurons. J Neurosci Res 2000; 61:364-370.    [Non-patent document 29] Iihoshi S, Honmou O, Houkin K et al. A therapeutic window for intravenous administration of autologous bone marrow after cerebral ischemia in adult rats. Brain Res 2004; 1007:1-9.    [Non-patent document 30] Honma T, Honmou O, Iihoshi S et al. Intravenous infusion of immortalized human mesenchymal stem cells protects against injury in a cerebral ischemia model in adult rat. Exp Neurol 2005 (in press).    [Non-patent document 31] Nomura T, Honmou O, Harada K et al. I.V. infusion of brain-derived neurotrophic factor gene-modified human mesenchymal stem cells protects against injury in a cerebral ischemia model in adult rat. Neuroscience 2005; 136:161-169.    [Non-patent document 32] Koc O N, Day J, Nieder M et al. Allogeneic mesenchymal stem cell infusion for treatment of metachromatic leukodystrophy (MLD) and Hurler syndrome (MPS-IH). Bone Marrow Transplant 2002; 30:215-222.    [Non-patent document 33] Koc O N, Gerson S L, Cooper B W et al. Rapid hematopoietic recovery after coinfusion of autologous-blood stem cells and culture-expanded marrow mesenchymal stem cells in advanced breast cancer patients receiving high-dose chemotherapy. J Clin Oncol 2000; 18:307-316.    [Non-patent document 34] Aggarwal S, Pittenger M F. Human mesenchymal stem cells modulate allogeneic immune cell responses. Blood 2005; 105:1815-1822.    [Non-patent document 35] Bang O Y, Lee J S, Lee P H et al. Autologous mesenchymal stem cell transplantation in stroke patients. Ann Neurol 2005; 57:874-882.    [Non-patent document 36] Zvaifler N J, Marinova-Mutafchieva L, Adams G et al. Mesenchymal precursor cells in the blood of normal individuals. Arthritis Res 2000; 2:477-488.    [Non-patent document 37] Huss R, Lange C, Weissinger E M et al. Evidence of peripheral blood-derived, plastic-adherent CD34 (−/low) hematopoietic stem cell clones with mesenchymal stem cell characteristics. Stem Cells 2000; 18:252-260.    [Non-patent document 38] Brown R A, Adkins D, Goodnough L T. Factors that influence the collection and engraftment of allogeneic peripheral-blood stem cells in patients with hematologic malignancies. J Clin Oncol 1997; 15:3067-3074.    [Non-patent document 39] Auner H W, Zebisch A, Ofner P et al. Evaluation of potential risk factors for early infectious complications after autologous peripheral blood stem cell transplantation in patients with lymphoproliferative diseases. Ann Hematol 2005; 84:532-537.    [Non-patent document 40] Bender J G, Unverzagt K L, Walker D E et al. Identification and comparison of CD34-positive cells and their subpopulations from normal peripheral blood and bone marrow using multicolor flow cytometry. Blood 1991; 77:2591-2596.    [Non-patent document 41] Tondreau T, Meuleman N, Delforge A et al. Mesenchymal stem cells derived from CD133-positive cells in mobilized peripheral blood and cord blood: proliferation, Oct4 expression, and plasticity. Stem Cells 2005; 23:1105-1112.    [Non-patent document 42] Rochefort G Y, Vaudin P, Bonnet N et al. Influence of hypoxia on the domiciliation of mesenchymal stem cells after infusion into rats: possibilities of targeting pulmonary artery remodeling via cells therapies?. Respir Res 2005; 6:125.    [Non-patent document 43] Chen J, Li Y, Wang L, et al. Therapeutic benefit of intracerebral transplantation of bone marrow stromal cells after cerebral ischemia in rats. J Neurol Sci 2001; 189: 49-57.    [Non-patent document 44] Chopp M, Zhang X H, Li Y, et al. Spinal cord injury in rat: treatment with bone marrow stromal cell transplantation. Neuroreport. 2000; 11(13):3001-3005.    [Non-patent document 45] Prockop D J. Marrow stromal cells as stem cells for nonhematopoietic tissues. Science 1997; 276:71-74.    [Non-patent document 46] Woodbury D, Schwarz E J, Prockop D J, et al. Adult rat and human bone marrow stromal cells differentiate into neurons. J Neurosci Res 2000; 61:364-370.    [Non-patent document 47] Kobune M, Kawano Y, Ito Y, et al. Telomerized human multipotent mesenchymal cells can differentiate into hematopoietic and cobblestone area-supporting cells. Exp Hematol 2003; 31:715-722.    [Non-patent document 48] Prockop D J, Gregory C A, Spees J L. One strategy for cell and gene therapy: Harnessing the power of adult stem cells to repair tissues. PNAS 2003; 100: 11917-11923.    [Non-patent document 49] Longa E Z, Weinstein P R, Carlson S, Cummins R. Reversible middle cerebral artery occlusion without craniectomy in rats. Stroke 1989; 20:84-91.    [Non-patent document 50] Nakamura Y, Wakimoto H, Abe J, et al. Adoptive immunotherapy with murine tumor-specific T lymphocytes engineered to secrete interleukin 2. Cancer Res 1994; 54:5757-5760.    [Non-patent document 51] Nakagawa I, Murakami M, Ijima K, et al. Persistent and secondary adenovirus-mediated hepatic gene expression using adenovirus vector containing CTLA4IgG Hum Gene Ther 1998; 9:1739-1745.    [Non-patent document 52] Takiguchi M, Murakami M, Nakagawa I, et al. CTLA4IgG gene delivery prevents autoantibody production and lupus nephritis in MRL/lpr mice. Life Sci 2000; 66:991-1001.    [Non-patent document 53] Neumann-Haefelin T, Kastrup A, de Crespigny A, et al. Serial MRI after transient focal cerebral ischemia in rats: dynamics of tissue injury, blood-brain barrier damage, and edema formation. Stroke 2000; 31:1965-1972.    [Non-patent document 54] Bederson J B, Pitts L H, Germano S M, Nishimura M C, et al. Evaluation of 2,3,5-triphenyltetrazolium chloride as a stain for detection and quantification of experimental cerebral infarction in rats. Stroke 1986; 17:1304-1308.    [Non-patent document 55] Villaron E M, Almeida J, Lopez-Holgado N, et al. Mesenchymal stem cells are present in peripheral blood and can engraft after allogeneic hematopoietic stem cell transplantation. Haematologica 2004; 89:1421-1427.    [Non-patent document 56] Willing A E, Vendrame M, Mallery J, et al. Mobilized peripheral blood cells administered intravenously produce functional recovery in stroke. Cell Transplant 2003; 12:449-454.    [Non-patent document 57] Hirouchi M, Ukai Y. Current state on development of neuroprotective agents for cerebral ischemia. Nippon Yakurigaku Zasshi 2002; 120:81-90.    [Non-patent document 58] Kurozumi K, Nakamura K, Tamiya T, et al. BDNF gene-modified mesenchymal stem cells promote functional recovery and reduce infarct size in the rat middle cerebral artery occlusion model. Mol Ther 2004; 9: 189-97.    [Non-patent document 59] Sasaki M, Hains B C, Lankford K L, et al. Protection of corticospinal tract neurons after dorsal spinal cord transection and engraftment of olfactory ensheathing cells. Glia. 2006; 53:352-359.    [Non-patent document 60] Chen X, Li Y. Wang L, et al. Ischemic rat brain extracts induce human marrow stromal cell growth factor production. Neuropathology 2002; 22:275-279.    [Non-patent document 61] Iuchino S, Zanotti L, Rossi B, et al. Neurosphere-derived multipotent precursors promote neuroprotection by an immunomodulatory mechanism. Nature 2005; 436: 266-271.    [Non-patent document 62] Keirstead H S, Ben-Hur T, Rogister B, et al. Polysialylated neural cell adhesion molecule-positive CNS precursors generate both oligodendrocytes and Schwann cells to remyelinate the CNS after transplantation. J. Neurosci 1999; 19:7529-7536.    [Non-patent document 63] Inoue M, Honmou O, Oka S, et al. Comparative analysis of remyelinating potential of focal and intravenous administration of autologous bone marrow cells into the rat demyelinated spinal cord. Glia 2003; 44: 111-118.    [Non-patent document 64] Sasaki M, Honmou O, Akiyama Y, et al. Transplantation of an acutely isolated bone marrow fraction repairs demyelinated adult rat spinal cord axons. Glia 2001; 35:26-34.    [Non-patent document 65] Hamano K, Li T S, Kobayashi T, et al. Angiogenesis induced by the implantation of self-bone marrow cells: a new material for therapeutic angiogenesis. Cell Transplant 2000; 9:439-443.    [Non-patent document 66] Bernstein D C, Shearer G M. Suppression of human cytotoxic T lymphocyte responses by adherent peripheral blood leukocytes. Ann NY Acad. Sci. 1988; 532:207-213.    [Non-patent document 67] Escolar M E, Poe M D, Provenzale J M et al. Transplantation of umbilical-cord blood in babies with infantile Krabbe's disease. The New England Journal of Medicine 2005; 352:2069-2081.    [Non-patent document 68] Staba S L, Escolar M L, Poe M, et al. Cord-blood transplants from unrelated donors in patients with Hurler's syndrome. N Engl J. Med. 2004; 350:1960-1969.    [Non-patent document 69] Zhang J, Li Y, Chen J, et al. Human bone marrow stromal cell treatment improves neurological functional recovery in EAE mice. Exp Neurol 2005; 195:16-26.    [Non-patent document 70] Pluchino S, Quattrini A, Brambilla E, et al. Injection of adult neurospheres induces recovery in a chronic model of multiple sclerosis. Nature 2003; 422:688-694.